CN104840625A - Liver-protecting pharmaceutical composition, and preparation method and use thereof - Google Patents
Liver-protecting pharmaceutical composition, and preparation method and use thereof Download PDFInfo
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Abstract
The invention provides a liver-protecting pharmaceutical composition. The liver-protecting pharmaceutical composition is a preparation prepared from the following medicinal raw materials in parts by weight: 24-36 parts of vine tea, 12-18 parts of pueraria flowers, 4-6 parts of Chinese olives, and 4.8-7.2 parts of liquorice. The invention further provides a preparation method and use of the pharmaceutical composition. The pharmaceutical composition disclosed by the invention is used for protecting livers and in particular has specific pharmacological effect to liver injury; after being dissolved in warm boiled water, a prepared effervescent tablet can be taken; the pharmaceutical composition is proper in mouth feel; the compliance of patients is increased; and therefore, the pharmaceutical composition has good market prospect.
Description
Technical field
The present invention relates to a kind of medicinal composition for protecting liver.Belong to drug world.
Background technology
In recent years, due to developing rapidly of society, the material life of people improves, and dietary structure is day by day enriched, people's life, operating pressure greatly, lack motion, often drink and stay up late, the sickness rate of hyperlipidemia, hepatitis rises year by year, brings very big inconvenience, jeopardize health to people's Working Life.Hyperlipidemia, hepatic injury day by day receive the concern of people, have dropped into the research and development that a large amount of manpower and materials carry out relevant medicine.Chinese medicine is the traditional medicine for disease preventing and treating of China, and the invaluable experience that to be people sum up in process in disease struggle, has a long history, aboundresources, plants, cost of gathering is low.But decocting for Chinese herbal medicine is loaded down with trivial details, carry inconvenience.
Ampelopsis grossedentata, being commonly called as Maoyanmei tea, Doanngo tea, rattan mother-in-law tea (also known as mountain Folium hydrangeae strigosae, dragon palpus tea), is the young stem and leaf of Vitaceae Ampelopsis Ampelopsis grossedentata section (Ampelopsis grossedentata) (Hand-Mazz) W.T.wang.Primary growth is in various places, China Chiang-Nan mountain area.This dark brown green bloom; The sweet length of micro-hardship, promoting the production of body fluid to quench thirst; Its sweet in the mouth sugariness is cool, has the effects such as heat-clearing and toxic substances removing, anti-inflammation, expelling wind and removing dampness, bone and muscle strengthening, blood pressure lowering, blood fat reducing, hepatoprotective.The control being usually used in the diseases such as hypertension, cold, fever, cardiovascular and cerebrovascular disease, dermatitis, eczema among the people.
After Ampelopsis grossedentata is soaked, tea bitterness is comparatively large, and is with puckery in hardship, and aftertaste is long, though have thereafter sweet, but the very large mouthfeel that have impact on user, and there is more deficiency in traditional product, urgently promotes in process of production.
Ampelopsis grossedentata medication history among the people was tracked to shennong went into the mountains collecting, tasting and testing different kinds of herbs to be used as medicine period, and " Book of Songs " is the earliest referred to as ancient tea Ramulus Uncariae Cum Uncis, and " tea warp " and " China's book on Chinese herbal medicine " also includes, and existing more than 2,000 year so far among the people drinks history.Fujian is produced Ampelopsis grossedentata and is had own characteristic: playing bloom, ripe perfume (or spice), flavour is abundant, elder generation is bitter, return sweet, promoting the production of body fluid to quench thirst afterwards, is the healthy tea-drinking of people from modern metropolitan cities.At present, except processing class tea product, Ampelopsis grossedentata also has the products such as beverage, buccal tablet, fine dried noodles, capsule, Chinese medicine formula tea and teabag, in a great variety.
Ampelopsis grossedentata conventional machining process is cut-out, parching to brown, oven dry, pulverizing, packaging, but this processing technique exists many deficiencies.One, Ampelopsis grossedentata all needs through processing, and Ampelopsis grossedentata total flavones and Destruction of Chlorophyll are comparatively serious in this course, and functional component and Color Quality lose more obvious; Its two, Ampelopsis grossedentata self exists that astringent taste is heavy, soup colour cast yellow and low vexed, the shortcoming that has foul smell of fragrance; Its three, current Ampelopsis grossedentata produces the production standard not having specification, and product is very different, and quality is unstable, and mechanization degree is not high, and production efficiency is low.Although processing class Ampelopsis grossedentata product category is various, the exploitation of Ampelopsis grossedentata resource are still on lower level generally.Substantially do not have according to the Ampelopsis grossedentata product of instruction of Chinese Medicine theory, product needs deeply to develop further, and the processing technique of product and standard also await setting up.
Application number: 97108163.8, denomination of invention: a kind of Ampelopsis grossedentata, the present invention relates to the health-care vine tea being refined acquisition by Folium Camelliae sinensis and Chinese medicine formula, it is characterized in that in Folium Camelliae sinensis, add the Chinese medicine formula being made up of different formulations and effect Ampelopsis grossedentata rattan and the Radix Astragali, Flos Chrysanthemi, Flos Lonicerae, Flos Jasmini Sambac, Radix Ginseng Rubra, flower of Radix Notoginseng.Its preparation method is: the medical material cleaning in Chinese medicine formula dried, and is placed in by weight ratio in volume to pulverize and puddles evenly and obtain Chinese medicine formula, finally puddled with ordinary tea leaves in proportion by Chinese medicine formula, toast drying.Disclosed in this patent, Ampelopsis grossedentata has the property of medicine of heat-clearing and toxic substances removing, preventing heatstroke, diuresis, also has the curative effect of tranquilizing by nourishing the heart and raising body immune system.
Summary of the invention
Technical scheme of the present invention there is provided a kind of medicinal composition for protecting liver.Another technical scheme of the present invention there is provided preparation method and the purposes of this pharmaceutical composition.
The invention provides a kind of medicinal composition for protecting liver, it is the preparation be prepared from by the crude drug of following weight proportioning:
Ampelopsis grossedentata 24-36 part, Flos puerariae lobatae 12-18 part, Fructus Canarii 4-6 part, Radix Glycyrrhizae 4.8-7.2 part.
Further preferably, it is the preparation be prepared from by the crude drug of following weight proportioning:
Ampelopsis grossedentata 30 parts, Flos puerariae lobatae 15 parts, Fructus Canarii 5 parts, 6 parts, Radix Glycyrrhizae.
Pharmaceutical composition of the present invention is active component by the protogenic medicinal powder of Ampelopsis grossedentata, Flos puerariae lobatae, Fructus Canarii, Radix Glycyrrhizae, water or extractive with organic solvent, adds the preparation that pharmaceutically acceptable adjuvant or complementary composition are prepared from.
Wherein, described preparation is tablet, pill, granule, capsule, oral liquid.
Wherein, described tablet is effervescent tablet.
Wherein, the weight percentage containing disintegrating agent in described effervescent tablet is 20-40%; In described disintegrating agent, sour material is the one in citric acid, tartaric acid; Alkali material is the one in sodium bicarbonate, sodium carbonate; The weight proportion of acid material and alkali material is: 1-1.2:1; Containing diluent in described effervescent tablet; Described diluent is the one in soluble starch, lactose, Icing Sugar; Containing binding agent in described effervescent tablet; Described binding agent is the one in ethanol, starch slurry, PVP; Containing lubricant in described effervescent tablet; Described lubricant is magnesium stearate, polyethylene glycol 6000; Weight percentage containing correctives in described effervescent tablet is: 0.45-1.8%; Described correctives is the one in sucrose, aspartame, stevioside, glycyrrhizin;
Further preferably, described disintegrating agent is tartaric acid, sodium bicarbonate; Described diluent is lactose; Described binding agent is PVP K30; Described lubricant is polyethylene glycol 6000; Described correctives is glycyrrhizin, and the weight proportion of supplementary material is:
Ampelopsis grossedentata 60 parts, Flos puerariae lobatae 30 parts, Fructus Canarii 10 parts, 12 parts, Radix Glycyrrhizae, 42.5 parts, tartaric acid, sodium bicarbonate 30.6 parts, lactose 247.0 parts, glycyrrhizin 8.5 parts, PEG60009.0 part, 5%PVP 0.4 part.
Present invention also offers the method for this pharmaceutical composition of preparation, it comprises the steps:
A, take the crude drug of each weight proportion;
B, Ampelopsis grossedentata, Flos puerariae lobatae, Fructus Canarii, Radix Glycyrrhizae decoct with water, and filter, filtrate is condensed into fluid extract, lets cool, and adds alcohol settling, leave standstill, get supernatant, reclaim ethanol, are condensed into thick paste; Add pharmaceutically acceptable adjuvant or complementary composition and be prepared into pharmaceutically conventional preparation.
Present invention also offers this pharmaceutical composition and prepare the purposes in hepatic.
Further preferably, described medicine is the medicine for the treatment of or/preventing liver injury.
Medicine material medicine of the present invention is made up of Ampelopsis grossedentata, Flos puerariae lobatae, Fructus Canarii, Radix Glycyrrhizae, can adjusting blood lipid, improves body immunity, has protective effect to chemical liver injury.Monarch drug Ampelopsis grossedentata is be the young stem and leaf of Vitaceae Vitis Ampelopsis grossedentata Ampelopsis grossedentata, and mainly containing effective constituent is dihydromyricetin, the effect such as it has resisting hypertension, blood fat reducing, protect the liver; Ministerial drug Flos puerariae lobatae, for the dried floral of leguminous plant Pueraria lobota Puerarialobata (Willd.) Ohwi, clinical trial shows its composition tectorigenin, iridin has strong liver-protecting activity, and kaikasaponin-III (KS-III) can reduce by I type enzyme hypoxanthine oxidase and Aldehyde oxidase activity plays reduction blood glucose and blood fat; Adjuvant drug Fructus Canarii, be the dry mature fruit of olive subject plant Fructus Canarii albi Canarium album Raeusch, the gallic acid etc. in its composition has hepatoprotective effect; Make medicine Radix Glycyrrhizae, the dry root and rhizome of glycyrrhizic legume Glycyrrhiza uralensis Fisch, the glycyrrhizic acid in composition has the effect of anti-liver injury, regulates immunity of organisms simultaneously, coordinating the actions of various ingredients in a prescription.
Medicine of the present invention is used for protecting the liver, especially hepatic injury, and drug effect is clear and definite, and the effervescent tablet be prepared into is taken after being dissolved in warm water, and mouthfeel is suitable for, and improves patient's compliance, has good market prospect.
Accompanying drawing explanation
Fig. 1 Normal group murine liver tissue HE × 200
Fig. 2 liver injury model group murine liver tissue HE × 200
Fig. 3 positive controls murine liver tissue HE × 200
Fig. 4 medicine effervescent tablet low dose group of the present invention murine liver tissue HE × 200
Fig. 5 medicine effervescent tablet high dose group of the present invention murine liver tissue HE × 200
Detailed description of the invention
The preparation of embodiment 1 effervescent tablet of the present invention
(1) prescription:
(2) preparation technology: Ampelopsis grossedentata, Flos puerariae lobatae, Fructus Canarii, Radix Glycyrrhizae are decocted with water three times, each 30 minutes, gradation filtered, and merging filtrate, is condensed into fluid extract, lets cool, and adds appropriate amount of ethanol, left standstill, got supernatant, reclaimed ethanol, was condensed into thick paste, for subsequent use.Add tartaric acid, lactose, glycyrrhizin, mixing, with 5%PVP alcoholic solution for binding agent, granulate, dry, granulate, adds PEG6000 and sodium bicarbonate, and mixing, is pressed into 1000, obtains final product.
Embodiment 2 extract drugs craft screening test of the present invention
1 instrument and reagent
(1) experimental apparatus
(2) reagent is tested
The Flos puerariae lobatae place of production: Beijing, Anhui three and pharmaceutcal corporation, Ltd
Ampelopsis grossedentata (place of production: Guangxi), Fructus Canarii (place of production: Guangdong), Radix Glycyrrhizae (place of production: Gansu): be all purchased from Foochow return of spring prepared slices of Chinese crude drugs company limited, all meet the Pharmacopoeia of the People's Republic of China (2010) year version relevant regulations through qualification.
The research of 2 extracting factor
2.1 orthogonal test screenings
Take Ampelopsis grossedentata, Flos puerariae lobatae, Fructus Canarii, each 0.5 times of recipe quantity of Radix Glycyrrhizae be 1 part, totally 9 parts, with amount of water (A), extraction time (B), extraction time (C) is investigation factor, carries out L9 (3 by the factor level of table 1
4) orthogonal test, evaluate according to general flavone content, yield of extract, measure its absorbance with ultraviolet spectrophotometer, investigate optimal extract process.
2.1.1 the mensuration of aqueous extract yield of extract
The every part of medicinal liquid 15ml extracted under drawing 2.1, transfers in dry evaporating dish, and be placed in water-bath after evaporation moisture content to fluid extract, put into baking oven and dry constant weight in 100 DEG C, weigh, record result also calculates.
2.1.2 the assay of aqueous extract total flavones
The preparation of reference substance solution: claim control substance of Rutin appropriate, accurately weighed, adds dissolve with methanol and is quantitatively diluted to the rutin solution of every 1ml containing 0.2mg, obtained reference substance solution.
The preparation of need testing solution: get above-mentioned 9 parts of each 1ml of extract, in 50ml volumetric flask, and be diluted with water to scale.
Prepared by control substance of Rutin standard curve: accurate absorption control substance of Rutin solution 0.0,1.0,2.0,3.0,4.0,5.0,6.0ml, puts 25ml (V respectively
3) in measuring bottle, add water to 6ml, add the 5%NaNO of 1ml
2, mixing, places 6min, adds the 10%Al (NO of 1ml in volumetric flask
3)
3, leave standstill 6min after mixing, add 10ml NaOH test solution, mixing, adding distil water is to scale, and mixing, leave standstill 15 minutes, the solvent prepared with 0.0ml reference substance solution is blank, measures the absorbance of each group under 510nm wavelength.Take absorbance as vertical coordinate (A), concentration is abscissa (mg/ml), drawing standard curve.
Sample determination: draw solution 10ml (V to be measured
2) in 25ml volumetric flask, according to the preparation of control substance of Rutin standard curve from " adding the 5%NaNO of 1ml
2" rise, the absorbance of working sample under wavelength 510nm, obtains the content (X) of total flavones in sample according to standard curve.
Result calculates:
Total flavones percentage composition in X-sample, with rutin (C
27h
30o
16) meter, g/100g (ml);
C-standard curve reads the concentration of total flavones in need testing solution, mg/ml;
V
1-sample constant volume, ml;
V
2-draw test liquid volume, ml;
V
3-colour developing constant volume, ml;
M-sample sampling amount, g (ml).
The results are shown in Table 2, table 3.
Table 1 extracts orthogonal test factor level table
Note: with D factor for error term
Adopt weighted mean method to carry out comprehensive assessment to yield of extract, general flavone content, because content is comparatively large on the impact of effervescent tablet curative effect, therefore, give yield of extract respectively, the weight of general flavone content 40%, 60% distributes.The rate of extractum is higher, and the dosage of the final single of patient is more, better lower, and with minimum yield 19.808% for top score, and total flavones is for mainly containing effective constituent, should be more high better, therefore with 1.969 for top score.Calculate comprehensive grading for sequence number 1: (19.808/19.808) * 0.4+ (0.796/1.969) * 0.6=0.643, in like manner, all the other each comprehensive gradings can be tried to achieve, the results are shown in Table 2.
Table 2 extracts orthogonal experiments (n=9)
Table 3 extracts orthogonal test variance analysis
As shown in Table 3, assess according to general flavone content in aqueous extract and yield of extract, each factor affects size to result: extraction time > amount of water > extraction time, namely major influence factors is extraction time, next is amount of water, and extraction time impact is minimum.Be reference with experimental result, in conjunction with practical condition, finally determine that extraction process is: add water 20 times amount, extracts 0.5h at every turn, extract 3 times.
2.2 extract orthogonal confirmatory experiment
According to above selected optimal extract process, add water 20 times amount, extracts 0.5h at every turn, extract 3 times, parallel 3 parts, the yield of extract of aqueous extract and general flavone content are measured, after calculating comprehensive grading by the method under 1.2.1.2 item, be averaging score, the results are shown in Table 4.
Table 4 extracts orthogonal experiment the result (n=3)
As shown in Table 4, the final average of 3 parts of parallel laboratory tests is 0.830, close to the A3B1C3 comprehensive grading in orthogonal design, and scoring is the highest, have good repeatability and higher accuracy rate, the extracting factor screened is basicly stable, can be used as the extracting method of medicine effervescent tablet of the present invention.
3 brief summaries and discussion
In the discussion process of extraction process, with the technique study amount of water of orthogonal test, extraction time, extraction time on the impact of medicinal liquid yield of extract and general flavone content, obtaining the suitableeest extraction conditions is: 20 times of water gagings, extracts 3 times, extracts 0.5h at every turn.Find in test, Fructus Canarii is the dry fruit of Fructus Canarii albi, and in this prescription, consumption is less, takes for convenience, improves the extraction ratio of the active ingredients such as gallic acid simultaneously, should first pulverize; Ampelopsis grossedentata, Flos puerariae lobatae density are little, bubble through the water column in leaching process, should by its abundant moistening, make it be immersed in water, in order to avoid affect the extraction ratio of each composition; If direct heating decocts, in leaching process, moisture evaporation is serious, and experimental result poor reproducibility, should adopt the method for condensing reflux to extract.
The research of embodiment 3 pharmaceutical purity metallization processes of the present invention
1 instrument and reagent
(1) experimental apparatus
All the other instruments are identical with under " 1.1 " item.
(2) reagent is tested
Dehydrated alcohol (AR) Chemical Reagent Co., Ltd., Sinopharm Group
All the other reagents used are identical with under " 1.1 " item.
The research of 2 effervescent tablet alcohol precipitation process conditions
2.1 precipitate with ethanol orthogonal test screenings
Take medical material Ampelopsis grossedentata, Flos puerariae lobatae, Fructus Canarii, each 0.5 times of recipe quantity of Radix Glycyrrhizae, totally 1 part, the optimum extraction process A3B1C3 screened by " 1.2.1 " item, namely 20 times of water gagings, extract 0.5h, extract 3 times, merge 3 extracting solution, be divided into 9 parts, every part of 350ml, with extract concentration (A), alcohol content (B), the precipitate with ethanol time (C) is investigation factor, carries out L9 (3 by the factor level of table 5
4) orthogonal test, and with total flavones extracted amount, dry extract yield for evaluation index, measure its absorbance with ultraviolet spectrophotometer, investigate best alcohol precipitation process condition.
2.1.1 the mensuration of precipitate with ethanol yield of extract
Precision measures medicinal liquid 5ml after above-mentioned precipitate with ethanol, measures yield of extract, the results are shown in Table 6 with reference to the method under " 2.1.1 " item.
2.1.2 the assay of total flavones after precipitate with ethanol
The preparation of reference substance solution: the preparation method with reference to the reference substance solution under " 1.2 " item is prepared.
The preparation of need testing solution: get each 1ml of medicinal liquid after above-mentioned 9 parts of precipitate with ethanol, put in 50ml volumetric flask, and be diluted with water to scale.
Prepared by control substance of Rutin standard curve: be prepared with reference to the control substance of Rutin standard curve preparation method under " 2.1.2 " item.
Sample determination: accurate absorption test solution 8ml (V
2), to in 25ml measuring bottle, according to " 2.1.2 " from " adding 5% sodium nitrite solution 1ml ", measure respectively at wavelength 510nm place and absorb angle value, the concentration (C) containing total flavones in need testing solution is read, the content (X) of total flavones in calculation sample from standard curve.
Computational methods: calculate with reference to the computational methods under " 1.2.1.2 " item.The results are shown in Table 6, table 7.
Table 5 precipitate with ethanol orthogonal test factor level table
Note: with D factor for error term
Adopt weighted mean method to carry out comprehensive grading to yield of extract, general flavone content, because content is comparatively large on the impact of effervescent tablet curative effect, therefore, give yield of extract respectively, the weight of general flavone content 40%, 60% distributes.The rate of extractum is higher, and the dosage of the final single of patient is more, should be more low better, therefore with minimum yield 18.68% for top score, and total flavones is for mainly containing effective constituent, should be more high better, therefore with 1.625 for top score.Calculate comprehensive grading for sequence number 1: (18.680/28.002) * 0.4+ (1.190/1.625) * 0.6=0.706, in like manner, all the other each comprehensive gradings can be tried to achieve, the results are shown in Table 6.
Table 6 precipitate with ethanol orthogonal experiments (n=9)
The variance analysis of table 7 precipitate with ethanol orthogonal test
As shown in Table 7, with general flavone content and aqueous extract yield of extract for evaluation index, the affect size of each factor on result is: relative density of medicine liquid > precipitate with ethanol time > alcohol content, namely major influence factors is relative density of medicine liquid, next is the precipitate with ethanol time, and alcohol content impact is minimum.Be reference with experimental result, in conjunction with practical condition, finally determine that alcohol precipitation process is: relative density of medicine liquid is 0.9959-0.9965, alcohol content 70%, precipitate with ethanol time 20h.
The orthogonal confirmatory experiment of 2.2 precipitate with ethanol
According to above selected best alcohol precipitation process condition, relative density is 0.9959-0.9965, alcohol content 70%, precipitate with ethanol time 20h, parallel 3 parts, the yield of extract of medicinal liquid after precipitate with ethanol and general flavone content are measured, after calculating comprehensive grading by the method under 2.1.2 item, be averaging score, the results are shown in Table 8.
Table 8 precipitate with ethanol orthogonal experiment the result (n=3)
As shown in Table 8, the average 0.918 of best alcohol precipitation process, is greater than the every comprehensive grading in orthogonal design, and scoring is the highest, and have higher accuracy rate, the alcohol precipitation process condition of screening is basicly stable, can be used as the alcohol precipitating method of medicine effervescent tablet of the present invention.
3 brief summaries and discussion
By orthogonal experiment, determine that best alcohol precipitation process is: be 0.9959-0.9965 by concentration of liquid medicine to relative density, alcohol content 70%, precipitate with ethanol time 20h.In precipitate with ethanol orthogonal test process, each experimental group adopts the extracting solution of same batch as far as possible, and to get rid of the impact of extraction factor on result, and extracting solution put procedure has a little precipitation, should first mix during sampling; When measuring relative density, survey after medicinal liquid to be concentrated to be cooled to room temperature, Yin Gaowen can be that the long-pending change of specific gravity of glass bottle is large, causes result inaccurate again; Add in the process of ethanol, the rate of addition of ethanol and mixing speed should be consistent, after the precipitate with ethanol time, isolate precipitate in time and reclaim ethanol with Rotary Evaporators.
Embodiment 4 medicine effervescent tablet of the present invention Study on Forming
1 instrument and reagent
(1) experimental apparatus
The grand experimental facilities company limited of Nereid on normal pressure thermostatic drying chamber XMTD-822
Single punch tablet machine TDP-1.5 Shanghai surpasses hundred million pharmaceutical machine equipment company limiteies
(2) reagent is tested
The screening of 2 moulding process prescriptions
The screening of 2.1 disintegrating agents and optimization
(1) the kind screening of disintegrating agent
Carry out soft material processed by the component in table 9, tabletting, disintegrate is tested, to screen disintegrating agent kind.
The preparation of table 9 variety classes disintegrating agent is investigated
Result shows, the effect of sample 4 is better.
(2) dosage optimization of disintegrating agent
In order to obtain the optimum amount of disintegrating agent, in table 9 based on sample 4, consult the consumption of corresponding document disintegrating agent mostly at 20%-40%, acid/base ratio is between 1:1-1.2:1, therefore design table 10, the consumption of different disintegrating agent and acid/base ratio, according to the disintegration rate of tablet, obtain optimum amount and acid/base ratio.
The consumption of table 10 disintegrating agent and acid/base ratio optimization
As seen from the above table, in disintegrating agent acid micro-excessive time disintegrate complete, better when ratio is 1.2:1; When the ratio of disintegrating agent in sheet is heavy increases, disintegration time reduces, but amplitude of variation is less, and when disintegrating agent ratio increases, the pH value of solution reduces, comparatively large to stomach injury, therefore selects disintegrating agent to account for sheet anharmonic ratio example 20%, and acid/base ratio is 1.2:1 is the most suitable.
The selection of 2.2 diluent
Should clarify because effervescent tablet dissolves solution later, therefore diluent should be able to be water-soluble, soluble starch, lactose, Icing Sugar are mainly considered in this experiment, because the extracting solution taste of medicine effervescent tablet of the present invention is more bitter, after adding Icing Sugar, extruding is granulated more difficult, and lactose easily sieves granulation, compressibility is strong, therefore selects lactose as diluent.
The selection of 2.3 binding agents
Conventional binding agent has ethanol, starch slurry, PVP.This experiment uses dehydrated alcohol, 10% starch slurry, 5% PVP K30 alcoholic solution as binding agent respectively, mixes respectively with sour material, alkali material, crosses 20 mesh sieve extruding and granulates, 60 DEG C of dry 30min, tabletting.Ethanol is as binding agent, and after dry, granule is looser, and segment more, mobility is poor, tablet weight variation is comparatively large, and starch slurry cohesive is poor, easily occurs loose pieces during tabletting, PVP K30 granule prevented from caking, disintegrate is good, therefore selects PVP K30 to make binding agent.
The selection of 2.4 lubricants
During wet granulation, usually need to add suitable lubricant before tabletting, to improve the mobility of granule, make sheet weight average even, tablet smooth in appearance.This experiment carries out contrast test to magnesium stearate, polyethylene glycol 6000, according to the size of angle of repose and bulk density, compares both impacts on mobility of particle.
2.4.1 the mensuration of angle of repose
Measure by fixing conical bottom method, funnel is fixed on iron stand, below place a filter paper, funnel lower end exit normal in its surface and aim at the center of circle, regulate highly about 4cm, the drug particles adding lubricant is poured into funnel from suitable for reading, until be paved with whole filter paper, measure the height of circular cone, calculate angle of repose: atg θ=h/r, parallel assay 3 times, the results are shown in Table 11.
2.4.2 the mensuration of bulk density
Get a 20ml graduated cylinder, cleaning, drying, weigh, granule is flowed down from about 5cm directly over bottleneck, until graduated cylinder endoparticle is accumulated to 20ml scale place, weighs graduated cylinder gross weight, try to achieve particle weight, parallel survey 3 times, average, according to weight and volume computing bulk density, the results are shown in Table 11.
Table 11 lubricant is investigated mobility of particle impact
From upper table bulk density, after adding lubricant, the stacking degree of powder body obviously increases, and namely the mobility of powder body, fillibility improve, and can reduce tablet weight variation in tableting processes, reduces the porosity between powder body simultaneously, strengthens compressibility; And angle of repose aspect, when not adding any lubricant, angle of repose is large, mobility is bad, and the coarse tarnish in tabletting rear surface, do not meet production requirement, after adding magnesium stearate, PEG6000, mobility is improved, tablet weight variation reduces, and tablet surface is smooth, but forms thin film in solution surface after magnesium stearate disintegrate, there is muddy sense, therefore using polyethylene glycol 6000 as lubricant.Find in experimentation, in the granule of identical weight, along with the increase adding lubricant, tablet surface polishes, but disintegration time also corresponding prolongation, experiment shows, when consumption is 2%, smooth surface, disintegrating property is good.
The selection of 2.5 correctivess
2.5.1 correctives kind is selected
Because the bitterness of medicine effervescent tablet extracting solution of the present invention is comparatively strong, suitable correctives need be adopted to correct.In this experiment, paper examines sucrose, aspartame, stevioside and the glycyrrhizin improvement situation to bitterness, result shows, sucrose due to sugariness little, in production process, addition is large, does not meet production requirement; Aspartame sugariness is comparatively large, but adds the hardening caking of rear medicine, is difficult to extruding and granulates; Stevioside, sugariness is large, takes rear astringent taste obvious, should not as correctives, and glycyrrhizin sugariness is large, and taking has back sweet between rear larynx, and contains Radix Glycyrrhizae in medicine effervescent tablet crude drug of the present invention, therefore selects glycyrrhizin as correctives.
2.5.2 correctives dosage optimization
Glycyrrhizin sugariness is larger, and individual sensory difference is comparatively large, therefore within the scope of correctives limitation, design addition is 0.45%, 0.90%, the medicine effervescent tablet of the present invention of 1.80% 3 gradient sugariness, carries out mouthfeel investigation, divides evaluate with 0-10 in crowd, mouthfeel satisfaction is better, score value is higher, is finally averaging score, obtains and accepts sugariness by masses.The results are shown in Table 12.
Table 12 different sugariness mouthfeel satisfaction investigation result (n=16)
As seen from the above table, when glycyrrhizin addition is less, the bitterness of drug extract is comparatively obvious, and increase with addition, bitterness is capped, and tartaric taste is obvious, and when addition is 1.8%, mouthfeel is satisfied.
Beneficial effect of the present invention is proved below by way of concrete pharmacodynamics test.
Test example 1 medicine effervescent tablet of the present invention is to the protective effect laboratory report of rat Acute Liver Injury Induced by carbon tetrachloride
1 experiment purpose
Observe medicine effervescent tablet of the present invention to the protective effect of rat Acute Liver Injury Induced by carbon tetrachloride.
2 experiment materials
2.1 experiment test medicine
Medicine effervescent tablet of the present invention (pharmaceutical college of Fujian University of Traditional Chinese Medicine self-control 0.45g/ sheet, prescription: Ampelopsis grossedentata 30g, Flos puerariae lobatae 15g, Fructus Canarii 5g, Radix Glycyrrhizae 6g.Function: have auxiliary protection function to chemical liver injury), adding distil water is mixed with the gavage liquid of variable concentrations.
2.2 experiment reagent
AST (AST) test kit, alanine aminotransferase (ALT) test kit, superoxide dismutase (SOD) test kit (Bioengineering Research Institute is built up in Nanjing), hydrochloric acid, glacial acetic acid etc. are traditional Chinese medicines group reagent.
2.3 experiment equipment refiners, spectrophotometer, centrifuge, electronic scale, mouse cage, gastric perfusion needle, analytical balance etc.
3 animal groupings and process
Cleaning grade ICR female mice, body weight 20-25g, Fujian University of Traditional Chinese Medicine's Experimental Animal Center provides.According to the basic demand of Ministry of Public Health " health food inspection and assessment technical specification ", laboratory animal is divided into five groups at random by body weight, often organizes at least 10 i.e. Normal group, liver injury model group, positive controls (grape pip procyanidin 30mg/kgBW), medicine effervescent tablet low dose group (20mg/kgBW) of the present invention, medicine effervescent tablet high dose group (40mg/kgBW) of the present invention.Experimental session is respectively organized its mouse oral gavage and is given tested material, and Normal group and liver injury model group such as to give at the capacity distilled water, continuous 30 days.Weigh a body weight weekly, according to body weight adjustment gavage amount.
In test the 31st day by fasting 16h overnight for each treated animal, liver injury model group and each tested material group mice, (amount to CCl by 5ml/kgBW
4dosage be 80mg/kgBW), gavage gives the 1%CCl diluted by vegetable oil
4, blank group gives to wait capacity vegetable oil.After giving carbon tetrachloride 24h, get blood through eyeball, separation of serum, measure Serum ALT, AST.Get liver, after weighing, calculate liver index.Get right lobe of liver, grind in homogenizer with the solution of ice-cold 150mmol/L KCl and make tissue homogenate, carry out the detection of liver MDA, liver SOD.Get leftlobe of liver, put in 10% formalin fixing, pending liver pathomorphology inspection and lesion score.
4 observation index and assay method
4.1 body weight and organ index
Weigh in respectively when experiment starts with at the end of experiment, to dissect after mice and weigh liver weight, calculating weight gain value and liver index with normal saline flushing liver immediately.
4.2 Serum ALT
Employing alanine aminotransferase test kit (improvement reitman-frankel method) measures the ALT in serum.
4.3 serum AST
Employing AST test kit (improvement reitman-frankel method) measures the AST in serum.
4.4 liver malonaldehyde (MDA) content
After sacrifice of animal, get its liver immediately, make the tissue homogenate of 10% (w/v) with 0.9% ice-cold normal saline, according to thiobarbituricacidα-(TBA) colorimetric method for determining MDA content in " Hygiene Toxicology ".
Preparation TCA-TBA-HCl mixed liquor: dense to 15gTCA, 0.375gTBA and 2.25ml HCI is dissolved in (50 DEG C of water-baths are dissolved) in 200ml distilled water, prepared before use.
4.5 liver superoxide dismutases (total SOD) are active
Make 1% (w/v) liver homogenate with 0.9% ice-cold normal saline, adopt superoxide dismutase test kit (xanthine oxidase) to measure the activity of superoxide dismutase.
4.6 liver pathomorphology inspections
Get liver lobus sinister 10% formalin to fix, do not do cross section and draw materials, paraffin embedding from leftlobe of liver, HE dyes.With 40 times of whole tissue slices of object lens Continuous Observation, observe the area in the visual field shared by various pathological changes in each visual field respectively and the pathological changes total score in the accumulative observed visual field.The lesion type of main detection has ballooning degeneration of liver cells (cell enlargement, endochylema is residual a little), the cohesion of steatosis (occurring in hepatocyte endochylema that the fat of distinct drips cavity), endochylema (endochylema strengthens addicted to Yihong), hepatocyte hydropic degeneration, hepatic necrosis (endochylema changes addicted to Yihong, coagulation necrosis) etc.
According to following standards of grading to each treated animal histopathological scores:
The Histopathological Features of table 13 ballooning degeneration of liver cells and score value
| Roughly normal | 0 point |
| The hepatocyte that balloon sample becomes accounts for 1/4 of the whole visual field | 1 point |
| The hepatocyte that balloon sample becomes accounts for 1/2 of the whole visual field | 2 points |
| The hepatocyte that balloon sample becomes accounts for 3/4 of the whole visual field | 3 points |
| The hepatocyte that balloon sample becomes accounts for the whole visual field | 4 points |
The Histopathological Features of table 14 hepatic cell fattydegeneration and score value
| Roughly normal | 0 point |
| The hepatocyte of steatosis accounts for 1/4 of the whole visual field | 1 point |
| The hepatocyte of steatosis accounts for 1/2 of the whole visual field | 2 points |
| The hepatocyte of steatosis accounts for 3/4 of the whole visual field | 3 points |
| The hepatocyte of steatosis accounts for the whole visual field | 4 points |
The Histopathological Features of table 15 hepatocyte hydropic degeneration and score value
| Have no the hepatocyte of hydropic degeneration | 0 point |
| The hepatocyte of hydropic degeneration accounts for 1/4 of the whole visual field | 1 point |
| The hepatocyte of hydropic degeneration accounts for 1/2 of the whole visual field | 2 points |
| The hepatocyte of hydropic degeneration accounts for 3/4 of the whole visual field | 3 points |
| The hepatocyte of hydropic degeneration accounts for the whole visual field | 4 points |
The Histopathological Features of table 16 hepatic necrosis and score value
| Have no non-viable non-apoptotic cell | 0 point |
| Be dispersed in that respective cells accounts for the whole visual field 1/4 | 1 point |
| Non-viable non-apoptotic cell accounts for 2/4 of the whole visual field | 2 points |
| Non-viable non-apoptotic cell accounts for 3/4 of the whole visual field | 3 points |
| The existence of non-viable non-apoptotic cell diffusivity accounts for the whole visual field | 4 points |
Between given the test agent any one dosage group and liver injury model group, in the liver cell lesions such as the change of balloon sample, steatosis, endochylema cohesion, hydropic degeneration or hepatic necrosis, hepatic necrosis degree alleviates, difference has significance, and other lesion type and liver injury model group is obvious alleviates or no significant difference, can judge that zoopery pathological examination is positive.
Between given the test agent any one dosage group and liver injury model group, the change of balloon sample, steatosis, endochylema cohesion, hydropic degeneration four kinds of liver cell lesion types and are increased the weight of and alleviate to exist simultaneously, difference has significance, and hepatic necrosis degree alleviates, difference has significance, then the score of its various pathological change can be added, hepatic necrosis is marked 2 times and is counted, carry out statistical analysis with total score, if difference has significance, can judge that zoopery pathological examination is positive.
5 statistical analysis
Data represent with mean ± standard deviation (x ± s), adopt SPSS13.0 statistical software, and application one factor analysis of variance compares group difference, and application LSD method compares between two.First carry out homogeneity test of variance by the program of variance analysis, variance is neat, calculates F value, F<F
0.05, conclusion: no significant difference between each group mean; F>F
0.05, P<0.05, carries out comparing between two between multiple sample average.
6 results judge
Meet the following conditions according to Ministry of Public Health " health food inspection and assessment technical specification ", can judge that given the test agent has assistant interventional effect to chemical liver injury: index any one of blood parameters ALT, AST two indexs is positive and histopathologic examination's result is positive.
7 experimental results
7.1 medicine effervescent tablets of the present invention are on the impact of Mouse Weight
Carry out variance analysis to the difference of weight gain during each group of mouse experiment, result display no difference of science of statistics (P>0.05), in table 17.Illustrate under prescribed conditions, tested material has no significant effect the weight of animals growth.The each group of mice mental status is good, has no untoward reaction.
Table 17 medicine effervescent tablet of the present invention is on the impact of Mouse Weight
7.2 medicine effervescent tablets of the present invention are on the impact of mice serum ALT and AST
From table 18, compare with Normal group, liver injury model group, positive controls, low dosage and high dose medicine effervescent tablet of the present invention treated animal Serum ALT levels obviously raises (P<0.01); The Serum ALT levels of positive controls, low dosage and high dose medicine effervescent tablet of the present invention group is all lower than liver injury model group (P<0.01); Serum ALT there was no significant difference (P>0.05) between medicine effervescent tablet group of the present invention; ALT level and the more equal not statistically significant of positive controls (P>0.05) of medicine effervescent tablet low dosage of the present invention and high dose group animal.
The AST level of liver injury model group, positive controls, low dosage and high dose medicine effervescent tablet of the present invention treated animal serum is all higher than Normal group (P<0.01); Positive controls, low dosage medicine of the present invention effervescent tablet group, high dose medicine effervescent tablet of the present invention group mice serum AST level are all lower than liver injury model group (P<0.01); Between two dosage medicine effervescent tablet of the present invention groups, mice serum AST is without significant difference (P>0.05); Serum AST there was no significant difference (P>0.05) compared with positive controls of two dosage medicine effervescent tablet of the present invention treated animals.
Table 18 medicine effervescent tablet of the present invention is on the impact of mice serum ALT and AST
A: compare with Normal group, P<0.01; B: compare with liver injury model group, P<0.01.
7.3 medicine effervescent tablets of the present invention are on the impact of mouse liver MDA content and SOD activity
From table 19, compare with Normal group, liver injury model treated animal liver MDA content obviously increases (P<0.01), and positive control and medicine effervescent tablet of the present invention each dosage treated animal liver MDA content are without significantly changing (P>0.05); The animal livers MDA content of positive controls and each dosage group of medicine effervescent tablet of the present invention is all lower than liver injury model group (P<0.01); There was no significant difference (P>0.05) between medicine effervescent tablet of the present invention each dosage treated animal liver MDA content, medicine effervescent tablet low dosage of the present invention and high dose group animal livers MDA content equal not statistically significant (P>0.05) compared with positive controls.
The SOD of liver injury model group, medicine effervescent tablet low dosage of the present invention and high dose group animal livers is active all lower than Normal group (P<0.01); The SOD activity of positive controls animal livers and the no significant difference (P>0.05) of Normal group; The SOD of positive controls, medicine effervescent tablet low dosage of the present invention and high dose group animal livers is active all higher than liver injury model group (P<0.01); Medicine low dose group animal livers SOD of the present invention is active in positive controls (P<0.05); SOD activity and positive controls difference remarkable (P>0.05) of medicine effervescent tablet high dose group animal livers of the present invention; The difference of the SOD activity of low dosage, high dose medicine effervescent tablet of the present invention treated animal liver is not significantly (P>0.05).
Table 19 medicine effervescent tablet of the present invention is on the impact of mouse liver MDA content and SOD activity
A: compare with Normal group, P<0.01; B: compare with liver injury model group, P<0.01;
C: compare with positive controls, P<0.05
7.4 medicine effervescent tablets of the present invention are on the impact of Mouse Liver body when liver tissue lesions's scoring
The liver index of liver injury model group mice is greater than Normal group (P<0.05); Positive controls and liver index with liver injury model group compared with the there was no significant difference (P>0.05) of medicine effervescent tablet low dosage of the present invention with high dose group animal.Difference not obvious (P>0.05) between each tested material treated animal liver index.Hepatocellular pathological changes integration is (with Hepatocellular ballooning, steatosis, endochylema condenses, hepatocyte hydropic degeneration, the total mark of five kinds of pathological changes such as hepatic necrosis is Judging index) display, liver injury model group, positive controls, low dosage and high dose medicine effervescent tablet of the present invention group mouse liver cell pathological changes integration are apparently higher than Normal group (P<0.01), positive controls and medicine effervescent tablet high dose group mouse liver cell pathological changes integration of the present invention are all lower than liver injury model group (P<0.05), medicine effervescent tablet low dose group mouse liver cell pathological changes integration of the present invention and liver injury model group difference are not significantly (P>0.05), there was no significant difference between each tested material group mouse liver cell pathological changes integration
(P>0.05)。In table 20.
Table 20 medicine effervescent tablet of the present invention is on the impact of Mouse Liver body when liver tissue lesions's scoring
A: compare with Normal group, P<0.05; C: compare with Normal group, P<0.01;
B: compare with liver injury model group, P<0.05
7.5 medicine effervescent tablets of the present invention are on the impact of mouse liver Histopathological Features
According to 2003 Nian Ban Ministry of Health of the People's Republic of China " health food inspection and assessment technical specification ", liver section carries out HE dyeing, result shows, normal group liver structure is complete, hepatocyte centered by central vein to surrounding radially proper alignment, lobules of liver clear-cut, hepatocyte boundary is clear, kytoplasm enriches, and has no pathologic and changes.Liver injury model group, liver normal organization disappears, and lobule boundary is unclear, and most of sinus hepaticus disappears.The hydropic degeneration of hepatocyte popularity, shows as cell volume and increases, endochylema is loose, understain, and even limpid, transparent, and part of hepatocytes is a small amount of stove or spotty necrosis.Positive controls and medicine effervescent tablet group pathological examination results of the present invention display lobules of liver sharpness of border, hepatocyte structure is without obvious degeneration, nuclear structure is more clear, part cell volume is bigger, caryoplasm is less slightly, or skewness, occur empty dizzy in part core, have the cohesion of ballooning degeneration of liver cells in various degree, hepatic cell fattydegeneration, liver cytoplasm, hydropic degeneration and hepatic necrosis.Show positive controls and medicine effervescent tablet of the present invention is low, high dose group has part improvement result to acute liver, positive control, high dose medicine of the present invention effervescent tablet, the impact of low dosage medicine of the present invention effervescent tablet on hepatic injury mouse liver are basically identical.See Fig. 1-5.
8 experiment conclusion
Medicine effervescent tablet of the present invention can alleviate increasing of ALT, AST level in the mice serum caused by carbon tetrachloride hepatic injury, reduce the generation of hepatic lipid peroxidation thing and/or accelerate the removing of peroxide, improve superoxide dismutase activity in hepatic tissue, improve the oxidation resistance of body, alleviate the hepatic tissue pathology damage caused by chemical poisonous substance.Thus, medicine effervescent tablet of the present invention can alleviate acute chemical hepatic injury, its effect may with antioxidation, scavenging free radicals, the structure of stabilizing cell membrane etc. effect relevant.
Claims (10)
1. a medicinal composition for protecting liver, is characterized in that: it is the preparation be prepared from by the crude drug of following weight proportioning:
Ampelopsis grossedentata 24-36 part, Flos puerariae lobatae 12-18 part, Fructus Canarii 4-6 part, Radix Glycyrrhizae 4.8-7.2 part.
2. pharmaceutical composition according to claim 1, is characterized in that: it is the preparation be prepared from by the crude drug of following weight proportioning:
Ampelopsis grossedentata 30 parts, Flos puerariae lobatae 15 parts, Fructus Canarii 5 parts, 6 parts, Radix Glycyrrhizae.
3. pharmaceutical composition according to claim 1 and 2, it is characterized in that: it is active component by the protogenic medicinal powder of Ampelopsis grossedentata, Flos puerariae lobatae, Fructus Canarii, Radix Glycyrrhizae, water or extractive with organic solvent, add the preparation that pharmaceutically acceptable adjuvant or complementary composition are prepared from.
4. pharmaceutical composition according to claim 3, is characterized in that: described preparation is tablet, pill, granule, capsule, oral liquid.
5. pharmaceutical composition according to claim 4, is characterized in that: described tablet is effervescent tablet.
6. pharmaceutical composition according to claim 5, is characterized in that: the weight percentage containing disintegrating agent in described effervescent tablet is 20-40%; In described disintegrating agent, sour material is the one in citric acid, tartaric acid; Alkali material is the one in sodium bicarbonate, sodium carbonate; The weight proportion of acid material and alkali material is: 1-1.2:1; Containing diluent in described effervescent tablet; Described diluent is the one in soluble starch, lactose, Icing Sugar; Containing binding agent in described effervescent tablet; Described binding agent is the one in ethanol, starch slurry, PVP; Containing lubricant in described effervescent tablet; Described lubricant is magnesium stearate, polyethylene glycol 6000; Weight percentage containing correctives in described effervescent tablet is: 0.45-1.8%; Described correctives is the one in sucrose, aspartame, stevioside, glycyrrhizin.
7. pharmaceutical composition according to claim 6, is characterized in that: described disintegrating agent is tartaric acid, sodium bicarbonate; Described diluent is lactose; Described binding agent is PVP K30; Described lubricant is polyethylene glycol 6000; Described correctives is glycyrrhizin, and the weight proportion of supplementary material is:
Ampelopsis grossedentata 60 parts, Flos puerariae lobatae 30 parts, Fructus Canarii 10 parts, 12 parts, Radix Glycyrrhizae, 42.5 parts, tartaric acid, sodium bicarbonate 30.6 parts, lactose 247.0 parts, glycyrrhizin 8.5 parts, PEG60009.0 part, 5%PVP 0.4 part.
8. prepare a method for the pharmaceutical composition described in claim 1-5 any one, it comprises the steps:
A, take the crude drug of each weight proportion;
B, Ampelopsis grossedentata, Flos puerariae lobatae, Fructus Canarii, Radix Glycyrrhizae decoct with water, and filter, filtrate is condensed into fluid extract, lets cool, and adds alcohol settling, leave standstill, get supernatant, reclaim ethanol, are condensed into thick paste; Add pharmaceutically acceptable adjuvant or complementary composition and be prepared into pharmaceutically conventional preparation.
9. the pharmaceutical composition described in claim 1-7 any one is preparing the purposes in hepatic.
10. purposes according to claim 9, is characterized in that: described medicine is the medicine for the treatment of or/preventing liver injury.
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| CN111184807A (en) * | 2018-11-14 | 2020-05-22 | 衡阳西汉至尊生物科技有限公司 | Traditional Chinese medicine composition for dispelling effects of alcohol and preparation method thereof |
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